1. Field of the Invention
This invention relates to a method and a kit for selecting and providing components or compounds to be used in formulating or compounding a drug which is to be used in a patient for cancer detection or therapy and quality control testing of the resulting compound.
2. Description of the Prior Art
The use of compositions which emit radiation at levels which can be detected after administration to a patient is well known in the art. The use of compositions which emit radiation at levels which can destroy cancerous tissues by concentrating the radioactivity in the area of tumors after the radiolabeled composition is administered to the patient is also well known in the art. However, the effective use of these compositions is limited when administered to patients with tumors that do not express the particular antigenic determinants recognized by the antibodies used in the composition. These prior art compositions rely on the selection of antibodies using the criteria of known or estimated prevalence of a particular antigen associated with a particular type of tumor. For example, colon tumors frequently express carcinoembryonic antigen (CEA), therefore an antibody composition for colon tumor is likely to contain antibodies reactive with CEA. Such an antibody, though, may not be the most effective choice for the treatment of a particular patient's tumor. Accordingly, it is highly desirable to determine whether a particular antibody binds to a given patient's tumor, in advance, in order that this antibody may be selected for inclusion in a drug or composition which is administered to the patient. It is also desirable to quality control a compound prior to administration to a patient to assure that the compound will bind to the tumor.
Prior art solid phase systems for determining reactions between antigens and antibodies have been widely used in a variety of assay systems, particularly radioimmunoassays. In such solid phase technology, the reagent or reagents used in the testing procedure are usually immobilized by being coated or bonded to a solid phase material such as a test tube or probe, which is then immersed in the sample to be tested. Additional reagents are then added to couple the solid phase material to an indicator. Commonly used indicators include the following: (1) antibodies coupled to enzymes and then further contacted with an enzyme substrate to give a color reaction; (2) antibodies coupled to a fluorescent substance which responds to certain frequencies of light or radiation exposure, and (3) antibodies coupled to radioisotopes which are detected by radiation detection devices or systems. Some solid phase systems employ lectins or other biological molecules which provide specific linkages between molecules. Examples of inventions which teach this type of assay include: U.S. Pat. No. 4,081,244, entitled "IMMUNOASSAY PROCEDURE EMPLOYING NOVEL IMMUNOCHEMICAL COMPOSITES", to Polito et al.; U.S. Pat. No. 4,092,408, entitled "METHOD FOR SOLID PHASE IMMUNOLOGICAL ASSAY OF ANTIGEN", to Litt; and U.S. Pat. No. 4,225,575, entitled "METHOD AND APPARATUS FOR PERFORMING IN VITRO CLINICAL DIAGNOSTIC TESTS USING A SOLID PHASE ASSAY SYSTEM", to Piasio et al. These prior art methods are useful for the detection and measurement of substances such as hormones or antibodies which occur in biological fluids such as sera. A common test using this prior art method is a pregnancy test in which the hormone hCG is detected in urine or blood. These prior art methods, however, are not useful in the detection of antibodies and antigens found in a tumor specimen nor do they provide means for selecting antibodies to be used in compounding a drug to be administered to an individual patient. In addition, these prior art methods are useful only in determining the presence and concentration of antigenic substances in solutions and not in determining which antibodies will react with a sample of a specific tumor. The present invention differs from the prior art in that the test material is derived from a solid tumor rather than from a liquid, and this tumor sample is fixed to the solid phase rather than reagents.
Methods for performing multiple simultaneous solid phase assays involving antigens and antibodies have been disclosed in the prior art. U.S. Pat. No. 4,378,344, entitled "METHOD AND APPARATUS FOR PERFORMING MULTIPLE, SIMULTANEOUS IN VITRO DIAGNOSTIC TESTS USING A SOLID PHASE SYSTEM," to Zahradnic et al.; teaches an assay method which utilizes a devise comprising a receptacle and an insert, each having solid phase reagents fixed to surfaces which are made to contact the fluid sample which is being analyzed. U.S. Pat. No. 4,459,360, entitled "MULTIPLE COMPONENT BINDING ASSAY SYSTEM AND METHOD OF MAKING AND USING IT," to Marinkovich, teaches an assay method which utilizes filaments in which each thread binds a different allergen. This assay system is particularity suited for the detection of IgE class antibodies in liquid samples. These prior art patents teach the use of solid phase reagents in the detection of different substances contained in common solutions. These prior art patents, however, are useful only for the testing of liquids, and specifically only for the testing of liquids which are used for in vitro diagnostic testing. The present invention, on the other hand, uses tumor tissue as the solid phase and tests it for reactivity with a series of different solutions of known compositions. This kind of testing cannot be achieved with prior art methods and devices.
Other prior art patents relating to the detection of antigens follow. U.S. Pat. No. 3,673,410 entitled "METHOD OF EXAMINATION OF CELL SAMPLES USING A RADIOACTIVELY TAGGED DYE," to Waite et al., teaches the screening of samples of tissue for the detection of disease. U.S. Patent No. 3,856,930, entitled "METHOD OF SCREENING TISSUE SPECIMENS FOR DIAGNOSTIC EXAMINATION," to Nodine et al., is an improvement over U.S. Pat. No. 3,673,410 by providing for the automatic elimination of specimens which do not need to be examined by more thorough diagnostic methods. Significant improvements over these approaches have resulted from methods which employ solid phase presentation of samples for the determination of one or more antigens or antibodies by detection of the corresponding antibody or antigen. U.S. Pat. No. 4,187,075, entitled "METHOD OF ANALYZING BIOLOGICAL, LIQUID SPECIMENS FOR ANTIGENS OR ANTIBODIES," to Noller, describes a method for detecting either an antigen or antibody in a solid phase presentation of a human body fluid. U.S. Pat. No. 4,495,295, entitled "IMMUNOASSAYS USING SUPPORT-CONTAINING SEPARATE ANTIGENS AND ANTIBODIES DERIVED FROM AN IMMUNE COMPLEX," to Neurath, improves upon previous methods for detecting antigens or antibodies by employing a dissociating buffer during the process to separate antigen-antibody complexes. U.S. Pat. No. 4,495,296, entitled "LABELED ANTI-HAPTEN ANTIBODIES AND THEIR USE AS A UNIVERSAL REAGENT FOR SOLID PHASE RADIO-AND/OR ENZYME-IMMUNOASSAYS," to Neurath et al., improves the detection of antigen and antibodies by employing anti-hapten antibodies as universal reagents. The present, invention improves on these previous inventions by using a panel of different antibodies in a parallel series which permit the detection of individual arrays of antigens to define the properties a given patient's tumor. The present invention provides a kit and method for matching antibodies to aliquots of tumor biopsy specimens, for selecting monoclonal antibodies to be used in compounding antibody-based drugs which are matched to the antigenic properties of a given tumor, and for quality control testing of the compounded drug.
U.S. Pat. No. 4,513,088, entitled "ASSAY FOR MONOCLONAL ANTIBODY AGAINST SURFACE IG OF A HUMAN B-CELL TUMOR," to Levy et al., describes a method for detecting antibodies by specifically screening hydridoma culture fluid for anti-idiotype antibodies. This method permits selection of antibodies useful in the treatment of human B-cell tumors. The present invention is an improvement over the Levy, et al., method because it permits selection of antibodies for the diagnosis and treatment of solid tumors. Furthermore, antibodies to general tumor associated antigens rather than to anti-idiotypic antibodies can be selected using the present invention.
The standard prior art method for determining if a specific monoclonal antibody will react with a tumor from an individual patient requires obtaining slices of the tumor, mounting the slices on a microscope slide, and staining the slice with the antibody in question. A separate microscope slide is required for each antibody when more than one antibody is being tested. An alternative prior art method is to take a sample of the tumor and treat it in such a way as to cause the individual cells of the tumor to disperse. The cells are reacted with antibodies labeled with fluorescent dye and passed through a cell sorter which counts the cells which have reacted with the antibody. Using this method, a separate cell sorter analysis is carried out for each antibody which is being tested. These prior art methods cannot be used for rapid, multi-antibody screening assays. Individual specimens have to be processed by specially trained individuals and tested one at a time for reactivity with a given antibody. Thus, these prior methods are costly, time consuming and difficult to standardize. The present invention provides for the simple and fast testing of a tumor specimen for reactivity with a panel of different antibodies, simultaneously, with quality control measures to ensure standardized testing.
A method for binding antigen to a solid phase for the detection of specific monoclonal antibodies has been reported by Suresh, M.R. and Milstein, C., Analytical Biochemistry, "A Direct Antigen--Binding Assay to Screen Hybridoma Supernatants." 151, 192-195 (1985). Using this method, the antigen is fixed to nitrocellulose paper spots which are attached a plastic insert. The insert is exposed to solutions which are to be assayed for the presence of antibody reactive with the fixed antigen. The presence of a specific antibody in a solution being tested is detected by exposing the nitrocellulose paper spot to a detecting solution comprising an anti-antibody conjugated to an enzyme which produces a color reaction when exposed to the appropriate enzyme substrate. This method is suitable for screening hybridomas to select those which secrete antibody which is specific for a particular antigen. This prior art method is used to find a particular antibody which may be present in the solution. This method, however, is not useful for determining which antigens are present and thus which antibodies or antibody fragments will react with a tumor specimen.
U.S. patent application Ser. No. 821,999, entitled METHOD AND APPARATUS FOR SELECTING ANTIBODIES OR ANTIBODY FRAGMENTS, discloses a method and apparatus selecting antibodies or antibody fragments for use in preparing drugs for cancer detection or therapy. The present invention is an improved method and kit for the selection, the compounding, and quality control testing of radiolabeled compounds and for the in vivo detection and treatment of cancer.
Use of prior art methods for the treatment or diagnosis of cancer often requires that a drug be administered to a patient first before determining whether or not that compound will have any beneficial effects. Even if a pre-treatment drug screening is performed for a patient, it is often too expensive to screen again. Prior art methods for screening cancer therapy drugs to predict the potential effectiveness of a given drug for use in an individual patient rely on growing tumor cells in tissue culture and exposing the growing cells to varying concentrations of different anticancer drugs. In vivo efficacy is predicted from the measured effect of the drug on the in vitro growth of the tumor cells in tissue culture. See "Applications of the Human Tumor Stem Cell Assay to New Drug Evaluation and Screening," by S.E. Salmon, Prog. Clin. Biol. Res, Vol 48, pp. 291-312, (1980). This prior art method is very laborious and can be used only in cases where the tumor cells can actually be grown in tissue cultures. The present invention avoids the necessity of tumor cell growth, and the necessity of measuring drug effects on tumor cell growth. An advantage of the present method is that when tumor cells undergo antigenic modification (the tumor changes its character) between treatments, the selection method of the invention can be used again to determine if different antibodies should be selected for the next treatment.
Accordingly, it is an object of the present invention to provide an inexpensive method and apparatus for selecting a panel of monoclonal antibodies which will specifically react with an individual patient's tumor.
It is a further object of the present invention to provide a method and apparatus for the simultaneous testing of several different monoclonal antibodies or antibody fragments.
Yet another object of the present invention is to provide a method and apparatus for the selection of antibodies in which a patient's tumor specimen is used as the solid phase reagent.
Another object of the present invention is to provide a kit and method for compounding antibodies or antibody fragments with a radionuclide for use in cancer detection or immunotherapy.
Another object of the present invention is to provide a method and apparatuses which contain quality control features for the standardization of antibody testing and for the testing of radiolabeled compounds or radiopharmaceuticals.
A further object of the present invention is to provide a method and an apparatus for optimizing the selection of antibodies, in advance, in the administration of drugs or compounds to an individual patient.
Other objects and further scope of applicability will become apparent from the detailed description to follow.